Development of loop-mediated isothermal amplification technique for LM canola T45, 73496, and MON88302 and its field application.

Autor: Choi, Wonkyun, Yoon, A-Mi, Lim, Hye Song, Lee, Jung Ro
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Zdroj: Plant Biotechnology Reports; Dec2023, Vol. 17 Issue 6, p947-954, 8p
Abstrakt: Brassica is an essential genus in agriculture, and gene flow from living modified (LM) organisms to native relatives or non-modified cultivars of the crucifer family is a critical issue in Asia. Loop-mediated isothermal amplification (LAMP) is a simple, rapid, and accurate method for identifying LM crops. This study aimed to develop a LAMP assay for the laboratory diagnosis and field analysis of three novel LM canola events: T45, 73496, and MON88302. The genomic DNA of each LM canola was amplified by incubating at 63 °C for 30 min with a newly developed 2 × LAMP premix containing Bst DNA polymerase and event-specific primer sets. The sensitivity of the LAMP assay for the three canola events in our condition ranged from 100 pg·μL−1 to 1 ng·μL−1. Genetic elements of the three LM events were identified through a combination of the LAMP assay, a syringe-type DNA extraction kit, and a portable isothermal amplifier. This study proposes a novel, simple system based on isothermal amplification for the DNA detection of the three LM canola events in a survey field. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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